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Microliter scale solid phase extraction devices

a solid phase extraction and microliter technology, applied in the field of solid phase extraction devices, can solve the problems of non-specific binding of analytes, difficult to achieve the effect of concentrating, purifying and separating analytes

Inactive Publication Date: 2006-07-27
SANDIA NAT LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0007] Provided herein are microliter scale solid phases extraction (μSPE) devices for microliter scale treatment of fluid samples. In one embodiment, the device is provided in a cartridge form that includes a central containment member having a containment bore disposed therethrough with openings at input and output ends of the containment bore. A volume defined by the containment bore between the input and output ends is less than 500 μl and more typically 1 to 10 μl or less. Each of the input and output ends of the first containment bore are surrounded by first and second sealing surfaces adjacent to the input and output ends, respectively. The cartridge further includes first and second conduit assemblies having a first and second conduit bores disposed therethrough with first and second openings, respectively, surrounded by a third and fourth sealing surfaces adjacent to the respective openings. A first porous membrane is sealingly engaged between the first sealing surface of the first containment bore and the third sealing surface of the first conduit assembly so that a perimeter of the first porous membrane is sealed from fluid flow while a first fluid contact area of the fir...

Problems solved by technology

Unfortunately, HPLC and CE systems are sophisticated and expensive instruments that require considerable sample processing (e.g., clean-up, fractionation and concentration) prior to actual analysis.
Retaining the chromatography media in the tips or columns in a stable position presents a problem.
Use of a frit, however, often introduces problems of non-specific binding of analytes as well as incomplete recovery and dilution of the sample.
Most importantly, these frits are fragile and can readily be broken or extruded.
A major problem with this solution is that the packed chromatography media can often escape the trapping matrix, making such devices unreliable for routine use.
Loose packing is particularly undesirable if the next step involves on-chip analysis or use of detectors, such as mass spectrometers, that can be damaged by these particles.
Yet another problem with all the above solutions is that it is difficult to control the rate and volume of fluid flow through the chromatography media to sub-microliter tolerances of about 1 μl or less.

Method used

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  • Microliter scale solid phase extraction devices
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  • Microliter scale solid phase extraction devices

Examples

Experimental program
Comparison scheme
Effect test

example 1

Concentration and Purification of BSA with a 5 μl μSPE Cartridge

[0060]FIG. 13 illustrates an example use of the μSPE cartridge 10 to concentrate and purify a sample protein. The containment bore 14 in cartridge 10 was packed with 5 μl of anion exchange beads containing DEAE (Toyo Pearl DEAE TSK from Tosoh Biosciences, Montgomeryville, Pa.) and tested for protein concentration. The cartridge was connected on one end to a syringe filled with 1 mM bovine serum albumin (BSA) in 5 mM borate buffer and to the other end to a fused silica capillary that was mounted in a UV detection setup. As the syringe delivers through the cartridge the BSA solution at a 10 μl / min flow rate, the absorbance of the solution that exits the cartridge is recorded by the UV detector. The breakthrough curve illustrates indirectly the amount of sample retained on the cartridge. As solution is passed through the cartridge part of the BSA is retained while the unretained BSA exits the cartridge and is recorded at ...

example 2

Protein Concentration Using 2 μl and 5 μl μSPE Cartridges Packed With Different DEAE or Hydrophobic Resins

[0062] 5 μl of a crude sample of BSA was injected in a microHPLC system to asses the amount of BSA present in the sample column. The lower U.V. absorbption chromatogram in FIG. 4A shows detection of a small peak of BSA at the expected position after 10 minutes of elution. 1 ml of the same crude sample was then treated by passing the entire contents through the first sample bore of a μSPE cartridge containing 2 μl of packed Toyo Pearl DEAE 650S and eluted with 200 mM NaCl into a 5 μl sample. The 5 μl sample was then subject to the same chromatographic analysis over the microHPLC reverse phase to generate the upper chromatographic trace. Comparison of the upper trace to the lower trace in FIG. 14A illustrates that the BSA was concentrated about 20 fold using the 2 μl SPE cartridge.

[0063]FIG. 14B illustrates concentration of a mixture of lactalbumin and lysozyme by treatment thou...

example 3

Reproducibility of the Same μSPE Cartridge

[0064] To test whether the μSPE cartridges provided herein are reusable and reliable over a period of time, the same procedure for concentrating and analyzing lactalbumin and lysozyme samples depicted in FIG. 14B and described in Example 2 was repeated using the same μSPE cartridge. Between uses, the μSPE cartridges were washed with 0.20 ml a washing buffer and reconditioned with 0.20 ml of the binding buffer. As illustrated in FIG. 15, the same μSPE cartridge showed reproducible recovery of about 70% of the input lactalbumin and about 60% of the input lysozyme.

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Abstract

Microliter scale solid phase extraction devices for preparing analytes in microliter volumes are disclosed. A re-useable device is provided in cartridge form that includes a central containment member with a containment bore that holds as little as 1 to 5 μl or less of a solid phase material that binds the analyte. The containment bore is enclosed on either side by a porous membrane that has an inner portion exposed for fluid flow and a peripheral portion that is sealed against fluid flow. The seal is formed by engaging the periphery of the porous membranes between sealing surfaces of the central containment member and corresponding sealing surfaces on first and second conduit assemblies that comprise the remainder of the cartridge. A non-reuseable device is provided in “chip” form, which includes a porous filter sandwiched between top and bottom wafers each having a plurality corresponding input and output conduits for a plurality of samples.

Description

STATEMENT REGARDING RESEARCH & DEVELOPMENT [0001] This invention was made with Government support under government contract no. DE-AC04-94AL85000 awarded by the U.S. Department of Energy to Sandia Corporation. The Government has certain rights in the invention, including a paid-up license and the right, in limited circumstances, to require the owner of any patent issuing in this invention to license others on reasonable terms.TECHNICAL FIELD [0002] This invention relates to solid phase extraction devices for extraction, concentration, separation and purification of analytes in microliter scale samples. BACKGROUND OF THE INVENTION [0003] Analytical instrumentation for biological and chemical analytes has increasingly become more sensitive and is now capable of detecting extraordinarily small amounts of sample materials that may be contained in microliter or submicroliter scale volumes. For example, micro HPLC systems are available for chromatographic separation of samples over column...

Claims

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Application Information

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IPC IPC(8): B01D63/00
CPCB01D61/18B01D61/28B01D63/08B01L3/502715B01L3/565B01L2200/027B01L2200/0631B01L2300/0681B01L2300/0816B01L2300/0877B01L2400/0421G01N1/405G01N30/02G01N2001/4016G01N2030/009B01D15/363B01D63/088B01D2313/22B01D2313/345B01D2313/50B01D2313/221
Inventor CHIRICA, GABRIELA S.RENZI, RONALD F.SIMMONS, BLAKE A.
Owner SANDIA NAT LAB
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